IMMUNOGENICITY ANALYSIS OF TRITERPENE GLYCOSIDE FROM HOLOTHURIA ATRA TO DETECTING FAS AND BCL-2 PROTEIN ON THE SP-C1 CELL OF TONGUE CARCINOMA

Objective: The objective of this study is to assess the role of triterpene glycoside of Holothuria atra to induce the Fas and Bcl-2-regulated apoptosis in
Supri’s Clone 1 (Sp-C1) cell of tongue carcinoma.
Methods: The triterpene glycoside of H. atra was isolated by high-performance liquid chromatography. The Sp-C1 cell of tongue carcinoma was cloned
by Dulbecco’s Modified Eagle Medium and cytotoxicity assay by 3-4-5-dimethylthiazol-2yl 2,5-diphenyltetrazolium bromide assay. Expression Fas and
Bcl-2 protein were analysed by immunocytochemistry also apoptosis detected by double staining ethidium bromide acridine. The datum of studied
was analyzed by one-way analysis of variance (ANOVA), significance (p<0.05), and strength correlation (p<0.001) with R=1.
Results: The H. atra has triterpene glycoside, and in the dose of 4 mg/ml, it has been cytotoxic activities on the Sp-C1 (p<0.05), mortality 80%;
inhibitory concentration 50 (IC50)=0.6 and anti-logarithm = 4. In general, the concentration of 2.5 mg/ml of triterpene glycoside has triggered the
expression of Fas protein (active, 71%; moderate, 10%; and no-active, 27%), whereas the Bcl-2 protein (active, 59%; moderate, 14%; and no-active
27%). Statistically, both expressions of protein were significant (p<0.05). Triterpene glycoside caused the apoptosis of Sp-C1 cell (strong, 87%, and
moderate, 13%).
Conclusion: The triterpene glycoside has the properties of cytotoxicity, and apoptosis in the SP-C1 cell also could be triggering the expression of Fas
and Bcl-2 proteins.

Keywords: Holothuria atra, Cytotoxic-apoptosis, Bcl-2 and Fas proteins, Supri’s Clone 1 cell, Triterpene glycoside.